Continuation is proposed of studies of the fine structures of heparins and heparan sulfates by fragmentation techniques. Isolation and characterization is being completed of a set of nonacetylated, sulfated and nonsulfated reference oligosaccharides (including di-, tri- and tetrasaccharides), which are produced by deamination of beef lung heparin and further chemical manipulation. The acetylated derivatives are also desired. Chemical synthesis of reference monosaccharide sulfates will be completed. Alternate fragmentation methods are also under study and will be particularly important for heparan sulfates. Mammalian glycosidases and sulfatases are being used as adjuncts to chemical methods. These include an apparently novel glucosamine-N,O-disulfate O-sulfatase. The methodology will be used to account for as much of the structure as possible of a preparation of beef lung heparin used as a model and source of fragments. The results obtained will be compared with similar results seen for other heparin and heparan sulfate preparations and with results obtained by others, using various working methods. The probable need for considerable difference in emphasis in studying heparan sulfates (which undergo only limited fragmentation with nitrous acid), is being kept closely in mind.